SCANNING ELECTRON MICROSCOPY OF THECAE OF THE DINOFLAGELLATE GENUS ORNITHOCERCUS1

Members of the genus Ornithocercus are all tropical oceanic species in which the theca is extended in the form of elaborate wing-like projections, the lists, supported by ribs. This paper illustrates the topography of 6 of the species. The theca is penetrated by numerous simple pores opening flush with the outer surface. On the inner side of the thecae the pores have a raised rim. The hypotheca of all species examined except O. splendidus have shallow depressions, the areolae, over most of the surface. Secondary thickening of mature cell walls deepens the appearance of the areolae, and increases their extent over the hypotheca in O. quadratus. The number of pores is not directly correlated with the areolae but seems rather to be a function of cell size. A comparison of the surface features confirms views that O. splendidus occupies a relatively isolated phenetic position. It also confirms the close similarity of O. steinii with O. thumii. Unexpectedly it suggested a phenetic closeness between O. magnificus and O. quadratus on the basis of hypothecal structure and rib features of the left sulcal list. O. heteroporus could not be subjected to the same degree of study and its position cannot be commented on. Some theoretical hydrodynamic and morphogenetic problems in Ornithocercus are discussed.     

CELL DIVISION IN THE DINOFLAGELLATE GAMBIERDISCUS TOXICUS IS PHASED TO THE DIURNAL CYCLE AND ACCOMPANIED BY ACTIVATION OF THE CELL CYCLE REGULATORY PROTEIN,CDC2 KINASE1

The cell division cycle in several pelagic dinoflagellate species has been shown to be phased with the diurnal cycle, suggesting that their cell cycle may be regulated by a circadian clock. In this study, we examined the cell cycle of an epibenthic dinoflagellate, Gambierdiscus toxicus Adachi and Fukuyo (Dinophyceae), and found that cell division was similarly phased to the diurnal cycle. Cell division occurred during a 3-h window beginning 6 h after the onset of the dark phase. Cell cycle progression in higher eukaryotes is regulated by a cell cycle regulatory protein complex consisting of cyclin and the cyclin-dependent kinase CDC2. In this report, we identified a CDC2-like kinase in G. toxicus that displays activity in vitro against a known substrate of CDC2 kinase, histone H1. As in higher eukaryotes, CDC2 kinase was expressed constitutively in G. toxicus throughout the cell cycle, but it was activated only late in the dark phase, concurrent with the presence of mitotic cells. These results indicate that cell division in G. toxicus is regulated by molecular controls similar to those found in higher eukaryotes.     

EFFECTS OF SMALL-SCALE TURBULENCE ON PHOTOSYNTHESIS,PIGMENTATION, CELL DIVISION,AND CELL SIZE IN THE MARINE DINOFLAGELLATE GOMAULAX POLYEDRA (DINOPHYCEAE)1

Several experiments were conducted to understand better the physiological mechanisms underlying growth inhibition of the dinoflagellate Gonyaulax polyedra Stein due to small-scale turbulence shear. To measure photosynthetic ¹⁴C uptake, a “phytoplankton wheel” device for rotating cultures in closed bottles was used. Turbulence was quantified biologically in the bottles by comparing growth inhibition with that in cultures with constant shear between a fixed cylinder and an outer concentric rotating cylinder (a stable Couette flow). At saturating irradiances, particulate photosynthesis (P_sat) or photosynthesis per unit chlorophyll (P^B_sat) were not inhibited completely at the highest turbulence level (26.6 rad.s^?1), and photosynthesis was less sensitive than growth. Photosynthesis per cell (P^C_sat) was increased by turbulence. In three experiments on the effects of turbulence on photosynthesis versus irradiance curves, the slope of the curve, α, for particulate photosynthesis at limiting irradiances did not change. Photosynthesis per unit chlorophyll per unit irradiance (α^B) decreased at high (but not intermediate) turbulence levels. Photosynthesis per cell per unit irradiance, α^C, increased with turbulence, suggesting an increase in photosynthetic efficiency in turbulent cultures. In two of the three experiments, respiration rates increased with turbulence, and in one experiment excretion of photosynthetically fixed ¹⁴C was not affected by motion. Ratios of accessory pigments to chlorophyll a did not change with turbulence, but pigments per cell and per dry weight increased with turbulence. These findings suggest little or no disruption of the photosynthetic apparatus. When turbulence was applied for 1 week, β-carotene increased while peridinin and diadinoxanthin decreased, suggesting inhibition of synthesis of these latter pigments by prolonged turbulence. Since cell numbers did not increase or decreased during turbulent 72–h incubations, cell division was inhibited and also the cells were very much enlarged. Increases in α^C per cell suggest that, in the sea, photo synthetic metabolism can persist efficiently without cell division during turbulent episodes. After turbulence ceases or reaches low levels again, cells can then divide and blooms may form. Thus, blooms can come or go fairly rapidly in the ocean depending on the degree of wave- and wind-induced turbulence.     

ULTRASTRUCTURE OF CELL DIVISION AND REPRODUCTIVE DIFFERENTIATION OF MALE PLANTS IN THE FLORIDEOPHYCEAE (RHODOPHYTA): CELL DIVISION IN POLYSIPHONIA1

Cell division in the marine red algae Polysiphonia harveyi Bailey and P. denudata (Dillwyn) Kutzing was studied with the electron microscope. Cells comprising the compact spermatangial branches of male plants were used exclusively because of their small size, large numbers and the ease with which the division planes can be predetermined. Some features characterizing mitosis in Polysiphonia confirm earlier electron microscope observations in Membranoptera, the only other florideophycean algae in which mitosis has been studied in detail. Common to both genera are a closed, fenestrated spindle, perinuclear endoplasmic reticulum, a typical metaphase plate arrangement of chromosomes, conspicuous, layered kinetochores, chromosomal and non-chromosomal microtubules, and nucleus associated organelles (NAOs) known as polar rings (PRs) located singly in large ribosome-free zones of exclusion at division poles in late prophase. However, other features, unreported in Membranoptera, were observed consistently in Polysiphonia. These include the presence of PR pairs in interphase-early prophase cells, the attachment of PRs to the nuclear envelope during all mitotic stages, the migration of a single PR to establish the division axis, a prominent, nuclear envelope protrusion (NEP) at both division poles at late prophase, the prometaphase splitting of PRs into proximal and distal portions, and the reformation of post-mitotic nuclei by the separation of an elongated interzonal nuclear midpiece at telophase. During cytokinesis, cleavage furrows impinge upon a central vacuolar region located between the two nuclei and eventually pit connections are formed in a manner basically similar to that reported for other red algae. Diagrammatic sequences of proposed PR behavior during mitosis are presented which can account for events known to occur during cell division in Polysiphonia. Mitosis is compared with that reported in several other lower plants and it is suggested that features of cell division are useful criteria to aid in the assessment of phylogenetic relationships of red algae.     

THE DINOFLAGELLATE PELLICULAR WALL LAYER AND ITS OCCURRENCE IN THE DIVISION PYRRHOPHYTA1

Forty-five species of dinoflagellates were surveyed for the presence of a pellicular layer in the amphiesma or cell covering. Such a layer was found in 15 of the 20 genera studied. Half the pellicles tested were resistant to acetolysis and may contain a sporopollenin-like material similar to that of some dinoflagellate cyst walk. Most organisms which formed pellicles were capable of reinforcing this layer with cellulose. Pellicles of Heterocapsa niei (Loeblich) Morrill & Loeblich and Scrippsiella trochoidea (Stein) Loeblich were studied with the electron microscope. Evidence is presented indicating that dividing cells of S. trochoidea from new walls while still enclosed in the parental pellicular layer.     

AN ULTRASTRUCTURAL STUDY OF CELL DIVISION IN THE CAMBIUM

The fine structure of dividing cambial cells of Ulmus americana and Tilia americana has been studied in material fixed in glutaraldehyde followed by osmium tetroxide. The cambia examined consisted of 7–9 rows of unexpanded fusiform cells, all of which had similar ultrastructural components. The fine structure and sequence of events of mitosis and cytokinesis in the dividing cambial cells apparently are similar to those of dividing cells in root tips and leaves. Of special interest was the observation that during cytokinesis, a broad cytoplasmic plate or phragmosome precedes the developing phragmoplast and cell plate through the dividing cambial cell. Smooth and coated vesicles derived from dictyosomes are associated with cell plate formation in these cells, smooth vesicles primarily with earlier stages of plate formation, and coated vesicles in later stages.     

cAMP IN THE CELL CYCLE OF THE DINOFLAGELLATE CRYPTHECODINIUM COHNII (DINOPHYTA)

The second messenger cAMP is a key regulator of growth in many cells. Previous studies showed that cAMP could reverse the growth inhibition of indoleamines in the dinoflagellate Crypthecodinium cohnii Biecheler. In the present study, we measured the level of intracellular cAMP during the cell cycle of C. cohnii . cAMP peaked during the G₁ phase and decreased to a minimum during S phase. Similarly, cAMP-dependent protein kinase activities peaked at both G₁ and G₂+M phases of the cell cycle, decreasing to a minimum at S phase. Addition of N6, O₂'-dibutyryl (Bt₂)-cAMP directly stimulated the growth of C. cohnii . Flow cytometric analysis of synchronized C. cohnii cells suggested that 1 mM cAMP shortened the cell cycle, probably at the exit from mitosis. The size of Bt₂-cAMP treated cells at G₁ was also larger than the control cells. The present study demonstrated a regulatory role of cAMP in the cell cycle progression in dinoflagellates.     

COMPLEX GENE STRUCTURE OF THE FORM II RUBISCO IN THE DINOFLAGELLATE PROROCENTRUM MINIMUM (DINOPHYCEAE)1

We analyzed the unusually complex organization of the nuclear‐encoded (form II) RUBISCO gene in the dinoflagellate Prorocentrum minimum (Parvillard) Schiller by intensive genomic DNA and cDNA sequencing and Western blotting. Over 10 transcribed units (TUs) were detected, which varied dramatically in their 3′ untranslated region. Each TU appeared to contain four tandem copies of the RUBISCO coding region (1.46 kb each; coding unit, or CU) interspersed by a 63‐bp spacer; the four CUs in each TU were cotranscribed and apparently cotranslated to a tetrameric polyprotein that may undergo successive cleavage steps to yield mature RUBISCO. By means of real‐time PCR analysis, it was estimated that each of the P. minimum genome harbored 148±16 CUs. Although nucleotide sequences varied by 1%–9% among the detected CUs, their inferred amino acid sequences were essentially identical. Our results suggest that the complex structure of Pmrbc has been derived from extensive and repeated gene duplications, an evolutionary process that has also been observed for other dinoflagellate genes.     

CELL DIVISION IN CHLAMYDOMONAS MOEWUSII1

Cell division in Chlamydomonas moewusii is described. The cells become immobile with flagellar abscission prior to mitosis. The basal bodies migrate toward the nucleus and become intimately associated with the nuclear membrane which is devoid, of ribosomes where adjacent to the basal bodies. The basal bodies replicate at preprophase. The nucleolus fragments at this stage. By prophase the basal body pairs have migrated, to the nuclear poles. Spindle fibers become prominent in the nucleus. The nuclear membrane does not fragment. The nucleus assumes a crescent-form by metaphase. Polar fenestrae are absent. Kinetochores appear at anaphase. An interzonal spindle elongates as the chromosomes move to the nuclear poles. Daughter nuclei become abscised by an ingrowth of nuclear membrane, leaving behind a separated, degenerating interzonal spindle. Ribosomes reappear on the outer nuclear membrane at late telophase. Nucleoli reform early in cytokinesis. The cleavage furrow, associated microtubules, and endoplasmic reticulum comprise the phycoplast. Cytokinesis proceeds rapidly after the completion of telophase. The basal body-nucleus relationship becomes reorganized into the typical interphase condition late in cytokinesis. Specific and predictable organelle rearrangements during mitosis have been described. Cell division in C. moewusii is compared with other algae, especially C. reinhardi.     

CELL DIVISION IN COSMARIUM BOTRYTIS1

Cell division in Cosmarium is described. Premitotic cells are very dense; the semicells, previously appressed to one another, separate slightly during entry into prophase. This separation coincides with deposition of a girdle of new wall material around the isthmus, where the 2 semicells are joined. Micro-tubules, abundant around the isthmus wall during interphase, all disappear during prophase; meanwhile, other microtubules proliferate outside the nuclear envelope. By metaphase, the nucleolus has dispersed, although remnants of it persist. The nuclear envelope breaks up, but some membranes coat metaphase chromosomes. The spindle, while being typical, is somewhat multipolar; microtubules, usually associated with elements of endoplasmic reticulum, are oriented toward numerous regions in the poles. During anaphase and telophase, these spindle tubules become increasingly directed toward a few discrete foci, and they persist after telophase. Meanwhile, the septum grows from the girdle of wall material to bisect the cell. After cytokinesis, some microtubules reappear near the isthmus, but only adjacent to the older, non-expanding semicell wall. Cell expansion then takes place, during which the nucleus, ensheathed in a complex microtubular system, moves into the forming semicell. Later, the chloroplast follows the nucleus and its 2 pyrenoids elongate and divide. When semicell expansion is complete, the chloroplast cleaves adjacent to the isthmus. The nucleus, now apparently not associated with microtubules, concurrently moves back into the isthmus. Continuous deposition of primary wall material accompanies cell expansion. Wall materials are apparently secreted as aggregates (perhaps derived from the contents of vesicles) adjacent to the plasmalemma, whose fibrous components become increasingly oriented in the outer layers of the wall by stretching. Late in semicell formation, this deposition ceases and during further expansion, the semi-cell develops a pattern of warts and ridges. Secondary wall deposition under the primary wall then follows, matching this pattern of ornamentation. In addition, numerous plugs of amorphous material capped by specialized regions of the plasmalemma, traverse the entire thickness of the secondary wall which becomes further thickened at these particular sites. The amorphous plugs presumably are eroded away later to form the mucilage pores of the vegetative cells. The wall microtubules gradually become more symmetrically arrayed around the isthmus as this new secondary wall thickens. These observations are discussed in comparison with other work on morphogenesis in desmids.     

THE FINE STRUCTURE OF MITOSIS AND CELL DIVISION IN THE CHRYSOPHYCEAN ALGA OCHROMONAS DANICA1

At the ultrastructural level, cell division in Ochromonas danica exhibits several unusual features. During interphase, the basal bodies of the 2 flagella replicate and the chloroplast divides by constriction between its 2 lobes. The rhizoplast, which is a fibrous striated root attached to the basal body of the long flagellum, extends under the Golgi body to the surface of the nucleus in interphase cells. During proprophase, the Golgi body replicates, apparently by division, and a daughter rhizoplast, appears. During prophase, the 2 pairs of flagellar basal bodies, each with their accompanying rhizoplast and Golgi body, begin to separate. Three or 4 flagella are already present at this stage. At the same time, there is a proliferation of microtubules outside the nuclear envelope. Gaps then appear in the nuclear envelope, admitting the microtubules into the nucleus, where they form a spindle. A unique feature of mitosis in O. danica is that the 2 rhizoplasts form the poles of the spindle, spindle microtubules inserting directly onto the rhizoplasts. Some of the spindle microtubules extend from pole to pole; others appear to attach to the chromosomes. Kinetochores, however, are not present. The nuclear envelope breaks down, except, in the regions adjacent, to the chloroplasts; chloroplast ER remains intact throughout mitosis. At late anaphase the chromosomes come to lie against part of the chloroplast ER. This segment of the chloroplast ER appears to be incorporated as part of the reforming nuclear envelope, thus reestablishing the characteristic nuclear envelope—chloroplast ER association of the interphase cell.     

CELL SIZE DIFFERENTIATION IN THE BLOOM-FORMING DINOFLAGELLATE GYMNODINIUM CF. NAGASAKIENSE1

Two subpopulations differing essentially by their mean cell size were observed regularly in cultures and natural samples of the naked dinoflagellate Gymnodinium cf. nagasakiense Takayama et Adachi (currently known as Gyrodinium aureolum Hulburt), a species which frequently forms red tides in North European seas. “Large” cells represented the typical forms; they were morphologically similar to cells of the closely related Japanese species G. nagasakiense, which did not form any subpopulation of reduced size. “Small” and “large” cells of G. cf. nagasakiense had the same DNA content, but the nucleus of the former appeared to be much more condensed during interphase. Each cell type was able to divide and had its own growth dynamics; therefore, any intermediary between pure populations of “small” and of “large” cells were observed in culture. The “large” form generated a “small” cell by an atypical budding-like division, whereas the “small” form gave back a “large” form, once it ceased to divide, by simple enlargement of its cell body. Factory inducing cell size differentiation are yet unclear. Neither nitrogen nor phosphorus starvation induced a significant increase in the relative proportion of “small” and budding cells. Although cell size differentiation is associated with the formation of gametes in a variety of dinoflagellates, we demonstrated that “small” cells of G. cf. nagasakiense are able to divide asexually, in contrast to gametes of most other species. The high proliferative power of “small” cells as compared with normal cells suggests that they could play a significant role during red tides of G. cf. nagasakiense; in contrast, cells of the Japanese species G. Nagasakiense could sustain high growth rates with larger cell size because this species generally blooms in waters much warmer than those found in northern Europe.     

图论在中国三尖杉属植物区系地理成分划分中的应用

本文根据图论中树与生成树的理论,选用中国三尖杉属(Cephalotaxus)8种植物的地理分布资料,采用图论分类法——Prim法,对国产三尖杉属植物地理分布进行定量研究。结果表明,本文应用图论分类法划分植物区系地理成分基本上是成功的,其结果与传统分析基本相符,尤其是采用不对称信息系数构成相异矩阵的方法;此外,本方法还有以下特点:(1)结果直观、简单;(2)能处理非常复杂的植物地理分布问题;(3)能避免因人为主观意识所引起的片面性甚至错误;(4)能准确地反映各类区系地理成分之间及其在地质时期植物地理联系的“亲缘关系”。     

ORIENTATION OF THE PLANE OF CELL DIVISION IN FERN GAMETOPHYTES: THE ROLES OF CELL SHAPE AND STRESS

Apical cells of Onoclea sensibilis L. protonemata were measured to determine areas of new walls which were formed during both transverse and longitudinal cell division. Actual wall areas were compared with calculated areas of hypothetical walls oriented in the opposite sense (i.e., an actual transverse wall compared with a hypothetical longitudinal wall, and the reverse). Among 87 out of 90 cells which were analyzed the actual walls had the least area. Thus, the minimal area hypothesis of cell partitioning accurately predicts wall orientation in this instance, although it appears, on other grounds, that the hypothesis does not furnish a plausible mechanism for wall orientation. The application of Lintilhac's concept of the orientation of cell walls in response to anisotropic stresses in the cell was explored. Photographs of apical cells during deplasmolysis indicated that unequal stresses might be generated in apical cells as a result of the osmotic distension of the elastic protoplast. It is concluded that the primary factor which determines the plane of cell division in the apical cell, and the transition from one- to two-dimensional growth, is the local pattern of stress which exists at the position of the nucleus at the time of onset of cell division and wall formation. Calculations of some geometrical properties of idealized model cells are interpreted to mean that the accuracy of the minimal area hypothesis results from a coincidence of its predictions with predictions of Lintilhac's hypothesis, and no causal significance is attributed to wall areas.     

VECTORIAL LABELING OF DINOFLAGELLATE CELL SURFACE PROTEINS1

Our objective was to identify cell surface proteins in the dinoflagellate Lingulodinium polyedrum. Proteins on the surface of living cells that had regions exposed to the external medium were labeled with Na¹²⁵I. After partial purification of membrane proteins and analysis by two‐dimensional gel electrophoresis and autoradiography, a protein of roughly 43 kDa was found to have incorporated the radiolabel. This protein was cloned using a combination of protein microsequencing and PCR amplification. The derived protein sequence in the cDNA has a signal peptide at the N‐terminal end of the protein and thus represents the first plasma membrane protein ever reported for a dinoflagellate. The function of the protein is unknown, but its cloning provides a proof of principle for the general use of vectorial labeling in identifying cell surface proteins of marine algae.